Synergistic Wound Healing Enhancement by Extracellular Vesicles From Engineered Macrophages and Fibroblasts.

Abstract

Severe wound healing impairment risks persistent tissue damage and even necrosis. Cell-free exosome therapy demonstrates significant potential in wound healing due to its high efficacy and micro-volume characteristics. However, its clinical-scale translation faces challenges including low yield, limited sources, functional monotony, and low bioavailability. We developed a lentiviral co-engineering strategy introducing exosome secretion-related genes into macrophage and fibroblast cell lines, achieving stable, high-yield (twofold increase) exosome production. Co-engineered extracellular vesicles (EVs) demonstrated enhanced cellular uptake and synergistically upregulated pro-angiogenic (HIF-1α, VEGF-A) and anti-inflammatory (IL-4, IL-10) factors. In a cell model and murine full-thickness wound model, the combination of EVs from both sources potently accelerated healing, promoting re-epithelialization and collagen deposition while orchestrating an anti-inflammatory response via TNF-α/IL-1β downregulation and IL-10 upregulation. In summary, co-cultured EVs not only exhibit high yield and purity but also demonstrate potent pro-angiogenic, anti-inflammatory, and antioxidant effects, effectively promoting wound healing, epithelial tissue regeneration, and collagen deposition. This study proposes an innovative and highly efficient method for synergistically regulating the production of EVs from cellular systems, providing promising research resources for their application in wound healing and medical esthetics. It also offers crucial insights for investigating the underlying mechanisms.